Laboratory of Signal Transduction, Department of Medical Sciences, Institute of Biomedicine, University of Aveiro, Aveiro, Portugal; Instituto de Investigação e Inovação em Saúde - i3S, University of Porto, Porto, Portugal; Laboratory of Cell Biology, Unit for Multidisciplinary Research in Biomedicine, Institute of Biomedical Sciences Abel Salazar, University of Porto, Porto, Portugal.
Laboratory of Signal Transduction, Department of Medical Sciences, Institute of Biomedicine, University of Aveiro, Aveiro, Portugal; present address: Laboratory of Protein Phosphorylation and Proteomics, Department of Cellular and Molecular Medicine, Faculty of Medicine, Catholic University of Leuven, Leuven, Belgium.
Fertil Steril. 2021 Feb;115(2):348-362. doi: 10.1016/j.fertnstert.2020.08.013. Epub 2020 Sep 23.
To design protein phosphatase 1 (PP1)-disrupting peptides covalently coupled to inert cell-penetrating peptides (CPPs) as sychnologically organized bioportide constructs as a strategy to modulate sperm motility.
Experimental study.
Academic research laboratory.
PATIENT(S)/ANIMAL(S): Normozoospermic men providing samples for routine analysis and Holstein Frisian bulls.
INTERVENTION(S): None.
MAIN OUTCOME MEASURE(S): Effect of the bioportides on the activity and interactions of PP1γ2-a PP1 isoform expressed exclusively in testicular germ cells and sperm-and on sperm vitality and motility.
RESULT(S): PP1-disrupting peptides were designed based on the sequences from: 1) a sperm-specific PP1 interactor (A kinase anchor protein 4); and 2) a PP1 inhibitor (protein phosphatase inhibitor 2). Those sequences were covalently coupled to inert CPPs as bioportide constructs, which were successfully delivered to the flagellum of sperm cells to induce a marked impact on PP1γ2 activity and sperm motility. Molecular modeling studies further facilitated the identification of an optimized PP1-binding sequence and enabled the development of a modified stop-sperm bioportide with reduced size and increased potency of action. In addition, a bioportide mimetic of the unique 22-amino acid C-terminus of PP1γ2 accumulated within spermatozoa to significantly reduce sperm motility and further define the PP1γ2-specific interactome.
CONCLUSION(S): These investigations demonstrate the utility of CPPs to deliver peptide sequences that target unique protein-protein interactions in spermatozoa to achieve a significant impact upon spermatozoa motility, a key prognostic indicator of male fertility.
设计共价连接到惰性穿透肽 (CPP) 的蛋白磷酸酶 1 (PP1) 破坏肽，作为一种调节精子运动的策略，将其构建为同步组织的生物肽。
实验研究。
学术研究实验室。
患者/动物：提供常规分析样本的正常精子症男性和荷斯坦弗里森公牛。
无。
生物肽对仅在睾丸生殖细胞和精子中表达的 PP1γ2-a PP1 同工酶的活性和相互作用，以及对精子活力和运动的影响。
根据以下序列设计 PP1 破坏肽：1）精子特异性 PP1 相互作用蛋白（A 激酶锚蛋白 4）；和 2）PP1 抑制剂（蛋白磷酸酶抑制剂 2）。这些序列被共价连接到惰性 CPP 上作为生物肽构建体，成功递送到精子细胞的鞭毛，对 PP1γ2 活性和精子运动产生显著影响。分子建模研究进一步促进了优化的 PP1 结合序列的鉴定，并开发了一种改良的停止精子生物肽，其尺寸减小，作用强度增加。此外，PP1γ2 独特的 22 个氨基酸 C 末端的生物肽模拟物在精子中积累，可显著降低精子运动能力，并进一步确定 PP1γ2 特异性相互作用组。
这些研究证明 CPP 可用于递送靶向精子中独特蛋白质-蛋白质相互作用的肽序列，从而对精子运动产生显著影响，精子运动是男性生育能力的一个关键预后指标。
